Look at molecules with a new twist of light
CHIRALIR-2X
TM
VCD Spectrometer
Characterization Experts: Chirality & Biologics
Vibrational Circular Dichroism
Chiral IR -2X VCD Spectrometer
V I B R AT I O N A L C I R C U L A R D I C H R O I S M ( V C D ) W A S F I R S T M E A S U R E D I N 1 9 7 3
Unique Advantages of the CHIRALIR-2X™ VCD Spectrometer
TM
In 1997, BioTools commercialized the technology
extends the functionality of electronic Circular
by introducing the ChiralIR™. We are proud to
Dichroism (CD) into the infrared spectral region
be the first company to present a stand-alone
where vibrational tranisitions in molecules are
dedicated VCD spectrometer and its powerful
observed.
applications in the fields of chemistry and
VCD combines the structural specificity of
biology.
Dual Source increases
signal-to-noise ratio (S/N) of approximately 3.6
times over any single-source system
FT-IR spectroscopy with the stereo-sensitivity of
30-48 hour hold detector time
circular dichroism. This gives access to multiple,
VCD is defined as the difference in the
Exclusive stage for elimination of
cell artifacts with SyncRoCell™
well-defined bands that provide molecular
absorbance (A) of left minus right circularly
quantitative information. Measurements can be
polarized infrared radiation, A=AL-AR. VCD
done in solids and solutions.
Simultaneous collection of IR & VCD
spectra (AC & DC)
Complete spectral range of
-1
measurement 4000-850 cm
(no change of filters required)
ONE MEASUREMENT GIVES TWO SPECTRA:
FT-IR AND VCD
A X 105
10
h
Sample spectrum in full range
(1S,2S)-( )-Pseudoephedrine
VCD
5
0
-5
A = AL - AR
Absorbance
1.0
H
IR
H3C
H3C N
H
0.5
H
OH
0.0
1500
1400
1300
1200
1100
Wavenumber (cm -1)
• Upgrade availability to patented DualPEM™ technology which allows artifact-free measurement of solids
E N A N T I O M E R I C PA I R S O F C H I R A L M O L E C U L E S
15
AX 105
10
• Factory aligned baseline resulting in immediate and routine operation
• Digital signal processing — no lock-in or electronic filter needed
(1R, 2R)-(+)-Pseudoephedrine
• Permanently aligned - Lifetime warranty on scan mechanism
5
• Training and support by the experts with unprecedented knowledge in design and applications of VCD
0
• Micro measurements with new VCDμSamplIR™
-5
-10
-15
1000
• ChiralIR-2X™ comes with standard IR cells (optional BioCell™ and TempCon available)
• ChiralIR-2X™ is also available in the Near-IR region.
(1S,2 S)-( )-Pseudoephedrine
1500
14001
-1
-1
The instrument can also be configured for coverage from 2000 to 4000 cm or from 4000 to 10000 cm
300
1200
Wavenumber (cm-1)
-2-
1100
1000
Standard IR Cell
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DualPEM Accessory (internal)
MANTIS DualPEM Accessory (external)
TM
TM
DualPEM™ provides significant improvement in baseline
position and stability over single PEM systems.
TM
MANTISTM ­— External DualPEM™ Option
The second PEM subtracts birefringence from optics in real-time
thus providing a significant reduction in artifacts, improved stability and is a
requirement for measuring VCD of solid samples.
• Now available as an internal or external accessory
2
• DualPEM™ is a patented technology of BioTools
1
DualPEM™inside the CHIRALIR-2X™ VCD Spectrometer
(R- S)/2
0
-2
Δ
A x10
4
-1 Dual Modulation
-3
Single Modulation
-4
ChiralIR-2X ™ Instrument
-5
-6
1650
1550
1450 1350 1250
1150
-1
W avenumber (cm )
1050
950
MANTISTM DualPEM™ Accessory
(connects seamlessly to side of ChiralIR-2X ™ )
External DualPEM™ Applications
• Reaction Monitoring
• Micro Measurements – VCDμSamplIR™
Sampling of solid samples at 1mm2 spatial resolution
• Magnetic VCD (MVCD)
• Matrix Isolation
Look at molecules with
a new twist of light
• Gas Sampling
...any other accessory that needs more space
than a standard sample or compartment
MANTISTM DualPEM™
To upgrade your existing FT-IR Spectrometer to VCD capability, please contact us
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-5-
VCD Accessories
Award Winning Patented Technology
ChiralIR-2X ™ Technology
• Thousands of molecules measured using the ChiralIR platform
SyncRoCell™
• Hundreds of patents on new compounds
Rotating stage for elimination of cell artifacts
• Hundreds of customer scientific papers using the ChiralIR series
• Used for submissions to regulatory agencies
• Articles in U.S. Pharmacopoeia
Te m p C o n ™
Award Winning Features & Technology
Te m p Con™ temperature controller unit is designed for
Peltier based temperature controller
use in FT-IR spectrometers with two types of windows:
large circular B i o Cell™ windows and rectangular
windows. The controller can be controlled either
manually or through computer input.
The Ramp-Soak Control Interface allows for
temperature studies that can be aligned with FT-IR
scanning software and can be used in applications
such as effect of temperature on protein conformation.
BioTools
U.S. Patents
W E P R O U D LY S E R V E H U N D R E D S O F C O M PA N I E S , U N I V E R S I T I E S A N D G O V E R N M E N T A G E N C I E S
AMGEN • Astellas • Astra-Zeneca • GlaxoSmithKline • Bristol-Myers Squibb
Eli Lilly • Janssen • Lupin • Merck • Pfizer • Takeda • and others...
W H AT O U R C U S T O M E R S A R E S AY I N G
BioCell™
The CaF2 cell for IR/UV-CD spectroscopy is created between a
perfectly flat, optically clear plate and another plate, the center of
which is deepened to form a recessed parallel surface surrounded by
a groove. The cells are very easily assembled and disassembled, filled
with solution, and washed between measurements.
“Over the last several years, the Vibrational Circular Dichroism (VCD) technique has dramatically revitalized the utility and
visibility of vibrational spectroscopy within pharmaceutical drug discovery. The ability of VCD to deliver absolute chiral
assignments without the need for standards or crystallizations, renders the technology a vital complement to traditional
pharmaceutical analytical technologies. “
Dr. Don Pivonka, AstraZeneca
“As a long-standing analytical chemist in pharmaceutical research with a focus on vibrational spectroscopy,
I recognized the potential impact VCD could have on many aspects of big pharma. With the ChiralIR I have
successfully assigned the configurations of more than 600 exploratory drug molecules, providing an international
service to research facilities in Europe and the United States.I consider this instrument to be world-class and a
bargain for the price given its state-of-the art technology and incredible level of reliability. “
Wide range of available path lengths:
~5-10, ~20, ~40, ~80 and ~120 mm
5 mm thickness x 50 mm diameter standard size.
Cells with 4 mm x 40 mm dimensions
and other custom sizes are also available
Dr. Douglas James Minick, Senior Research Investigator, GlaxoSmithKline
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-7-
Software
ComputeVOA™ Software
An all inclusive package for calculation of VCD spectra. Combines:
structure building, extensive conformational search, easy integration
with Gaussian09 and plotting of calculated spectra.
CompareVOA™ Software
Confidence level algorithm for comparing VCD (and ROA) experimental and
theoretical spectra; the output generates two plots: IR and VCD comparisons
of measured and calculated spectra, and statistical plot against a database of
prior comparisons.
Screen Shot Coming
ComputeVOA orchestrates a complex molecular dance. Data enters the stream through a molecule builder/
structure importer then moves into a robust molecular mechanics conformational search algorithm. The results
then pass through a transparent handshake to Gaussian 09 (Gaussian, Inc. Pittsburgh,PA) where quantum
mechanics determines the molecule’s structure and properties. It then returns seamlessly to ComputeVOA for
a Boltzmann-averaging of conformers and plotting of the IR and VCD spectra. The final output: The specific
stereo isomers’ 3D structure and accompanying InfraRed and Vibrational Circular Dichroism spectra.
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ChiralIR-2X ™ Software
Included with the CHIRALIR-2XTM. Easy to use, no learning curve.
-9-
Application Examples
Chiral Drugs
Chiral Drugs Quality Control
Since commercial introduction of VCD by BioTools thousands of compounds have been determined using
VCD can be used as a chiral measure for characterization of raw materials for process analytical
technology (PAT) during development, synthesis, formulation, and final production of drug
substances and drug products. VCD also can be used to test the interaction between formulated
chiral drug substances and excipients.
this technology. It has become the-must-have-tool in the pharmaceutical and other industries.
A B S O L U T E C O N F I G U R AT I O N D E T E R M I N AT I O N S
Step 1: Choose Configuration
e.g. (R) - Mirtazapine Configuration
Step 3: measure IR (bottom) and VCD (top) spectra
Step 2: Calculate IR (bottom) and VCD
of available enantiomer, e.g. (-) -Mirtazapine
(top) spectra of (R) - Mirtazapine
1.0
200
A
0.5
100
H3C
N
H
N
N
“The absolute configuration of chiral compounds is critical in understanding structure-activity relationships,
and in developing appropriate chiral separation, resolutions, or synthesis.”
“Vibrational Circular Dichroism (VCD) data are now being used to support New Drug Applications and
patents.”
Comparison of observed VCD spectra (right) with calculated VCD spectra
(left) unambiguously determines absolute configuration (-)-(R)-Mirtazapine and
conformation in solution as shown above.
“The bottom line with VCD or any new analytical method is if the data are convincing to the scientists
developing the drug, then that data should be convincing to the scientists here at the FDA.”
Stephen P. Miller, Office of New Drug Chemistry,FDA’s Center for Drug Evaluations & Research
Chemical & Engineering News - 7/18/05
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VCD Application Examples
Proteins - Peptides - Carbohydrates - Nucleic Acids
Reaction Monitoring
PROTEIN / PEPTIDE STRUCTURAL STUDIES
ASYMMETRIC REACTION MONITORING
b) Characterization of Fibrils
a) Secondary Structure
AX 105
VCD SHOWS A HIGH SENSITIVITY TO
24
G
20
F
16
E
12
D
8
C
4
B
0
A
-4
STRUCTURE OF FIBRILS
H
+
O
O
H
C
CF3COOH
OH
+
CF3COOH
O
OH
HO
H
O
HO
O
H
+
H
100
VCD
IR
1.2
CCl4
MCH
80
G: Concanavalin A
F: -Chymotrypsin
Formative Stages of Amyloid Fibrils
60
0.8
D: Lysozyme
40
C: Cytochrome c
0.4
Predicted EE%
Absorbance
E: Ovalbumin
B: BSA
20
A: Myoglobin
0.0
1800
1600
Wavenumber (cm -1)
1400
Proto-Filament/
Filament
Proto-Fibril/
Fibril
Mature Fibril
VCD provides an enhanced sensitivity to secondary structure of proteins and peptides. The changes in VCD spectra
are observed in both the Amide I and II regions. The VCD sign of Amide I band changes for proteins with the two
most common motifs - α-helical protein myoglobin (lower spectra) and a primarily
β-sheet containing proteins concanavalin A and chymotrypsin
(upper spectra).
F E AT U R E S :
0
02
02
04
04
06
06
Time (hr)
During the course of a chemical stereospecific reaction, the combination of IR, for detection of changes
in mole fraction composition, and VCD, for combined mole fraction composition and enantiomeric
excess (%EE), enables one to follow %EE of each chiral species as a function of time. The figure
above shows epimerization versus time of DDM with trifluoroacetic acid in two different solvents.
• The largest FT-VCD protein database available
• Dedicated software for protein data analysis
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080
- 13 -
Lab Services
Customer Support
CHIRALIR-2X ™ Installation
• Installation and training on measurement and interpretation of VCD spectroscopy of small molecules
by experienced scientists
CHIRAL MOLECULES
• Absolute Configuration
• VCD Measurements
• Qualification provided (IQ/OQ/PQ)
• ab initio calculations (VCD, ECD, ROA)
• BioTools’ combined service and application knowledge
• VCD Consulting & Training
Technical Support and Maintenance
BioTools provides users of CHIRALIR-2XTM with complete maintenance and technical support. Each
instrument sold comes with a full one year warranty. Additional maintenance contracts can be purchased.
• Chiral Methods Development
PROTEINS / PEPTIDES
• % Enantiomeric Excess
• Secondary Structure
• Small Scale (~1g) Purifications
E-mail based help desk is open 365/24/7 at [email protected]
• Formulation of Biologics
Consulting
• Effect of Environmental Changes - temperature, pH, storage
BioTools offers consulting services that bring our expertise on spectroscopy of chiral molecules to bear on
your applications and needs. Costs depend on the extent of the study and degree of urgency.
• Structural comparison of production lots
• Comparability
Vibrational Optical Activity: Principles and Applications
by Laurence A. Nafie
This unique book stands as the only comprehensive introduction to vibrational optical activity
(VOA) and is the first single book that serves as a complete reference for this relatively new,
but increasingly important area of molecular spectroscopy.
“Vibrational Optical Activity: Principles and Applications” will take the reader from the basic theory
through the practical and instrumental approaches, providing a unified, comprehensive
description to the field of VOA that gives both introductory and in-depth coverage to VCD
and ROA. Applications include the analysis of all classes of chiral molecules, including organic
and inorganic molecules, metal complexes, pharmaceutical and natural product molecules, and
the full range of biological molecules such as amino acids, peptides, sugars, proteins, protein
fibrils, carbohydrates, nucleic acids, viruses and bacteria.
Written in a thorough and progressive style, it will appeal to advanced undergraduates, graduates and research groups
in academia as well as researchers and technicians in the pharmaceutical and biotechnology industries.
Signed Copies Available
ChiralIR-2X ™
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- 15 -
Characterization Experts: Chirality & Biologics
Prota-3STM FT-IR
ChiralRAMAN-2XTM (ROA, Now with AFM / TERS)
µBioRAMANTM
MantisTM (DualPEM VCD accessory)
ComputeVOATM
CompareVOATM
BioTools Worldwide
Corporate Headquarters
North America
17546 Bee Line Highway
Jupiter, Florida 33458 USA
Phone: 561.625.0133
Fax: 561.625.0717
[email protected] • www.btools.com
Europe • Asia • Middle East • South America
IF-720-MCI-0001 • REV. 2.0 • 092014